肉牛源毛孢子菌Trichosporon loubieri的分离鉴定与药敏试验

doi:10.3969/j.issn.1004-1524.2018.01.04

浙江农业学报 ›› 2018, Vol. 30 ›› Issue (1): 26-35.DOI: 10.3969/j.issn.1004-1524.2018.01.04

肉牛源毛孢子菌Trichosporon loubieri的分离鉴定与药敏试验

钟锐¹, 姜尧章¹, 马晓平^{1, *}, 左之才¹, 黄小丽², 邓俊良¹, 沈留红¹, 余树民¹

1.四川农业大学动物医学院,四川成都611130;
2.四川农业大学动物科技学院,四川成都 611130

收稿日期:2017-07-31 出版日期:2018-01-20 发布日期:2018-02-09
通讯作者: 马晓平,E-mail: mxp886@sina.com.cn
作者简介:钟锐(1996—),女,四川成都人,本科生。E-mail: 1194592501@qq.com
基金资助:
四川农业大学科研兴趣计划项目(04051750); 现代农业产业技术体系四川省肉牛创新团队

Isolation, identification and drug sensitive test of Trichosporon loubieri from beef cattle

ZHONG Rui¹, JIANG Yaozhang¹, MA Xiaoping^{1, *}, ZUO Zhicai¹, HUANG Xiaoli², DENG Junliang¹, SHEN Liuhong¹, YU Shumin¹

1. College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China;
2. College of Animal Science and Technology,Sichuan Agricultural University, Chengdu 611130, China

Received:2017-07-31 Online:2018-01-20 Published:2018-02-09

摘要/Abstract

摘要： 为探究肉牛真菌性皮肤病的病因并有效防治该病。取发生皮癣病肉牛患部的皮屑、被毛进行真菌学检查和分离。采用沙保弱氏培养法对皮屑和毛发样品进行分离纯化。对分离株进行形态学、分子生物学鉴定和小鼠致病性、药敏试验。最终从样本中分离到一株真菌(编号zrfg01)。对分离株进行PCR扩增,获得长度为542 bp的转录基因间隔区(intergenic spacer,IGS)序列片段,序列提交GenBank,获得序列号为MF401393.1。序列在Blast中比对结果显示,zrfg01与编号KT936593.1的Trichosporon loubieri的相似性为100%。根据形态学和分子生物学方法将其鉴定为T. loubieri。动物实验表明该菌对小鼠有选择性致病作用。药敏试验表明,该菌在25 ℃环境条件下,对特比萘芬和氟胞嘧啶耐药,对克霉唑、两性霉素B、氟康唑、伏立康唑较为敏感。该研究可为诊断和治疗T. loubieri引起的肉牛皮肤癣菌病提供重要参考。

关键词: Trichosporon loubieri, 形态学, ITS序列, IGS序列, 致病性, 药敏试验

Abstract: In order to have a deeper understanding of the causes of skin disease that beef cattle suffered and find the effective treatment, we collected the dandruff and hair of beef cattle skin with dermatophytosis then carried our an inspection and a separation of mycology. The dandruff and hair of beef cattle skin with dermatophytosis were collected and used for fungi culture. The morphological characteristics and IGS sequences of the isolated strains were identified. The pathogenicity test was carried out in mice, and drug sensitive test was performed. Finally, one strain of fungi, zrfg01, was obtained. The length of the IGS sequence was 542 bp amplified by PCR, which was submitted to GenBank (ID, MF401393.1). NCBI blast result showed that zrfg01 had 100% similarity with Trichosporon loubieri. According to the morphological identification, IGS sequence analysis and phylogenetic tree, the isolated strain was identified as T. loubieri. The pathogenicity test revealed that the fungus had a selective pathogenesis to mice. The drug susceptibility test showed that under the environment of 25 ℃, T. loubieri was resistant to terbinafine and fluorouracil, but sensitive to clotrimazole, amphotericin B, fluconazole and voriconazole. This study provided an important reference for the diagnosis and treatment of dermomycosis caused by T. loubieri.

Key words: Trichosporon loubieri, morphology, ITS sequence, IGS sequence, pathogenicity, drug sensitivity.

中图分类号:

S858.23
R379

钟锐, 姜尧章, 马晓平, 左之才, 黄小丽, 邓俊良, 沈留红, 余树民. 肉牛源毛孢子菌Trichosporon loubieri的分离鉴定与药敏试验[J]. 浙江农业学报, 2018, 30(1): 26-35.

ZHONG Rui, JIANG Yaozhang, MA Xiaoping, ZUO Zhicai, HUANG Xiaoli, DENG Junliang, SHEN Liuhong, YU Shumin. Isolation, identification and drug sensitive test of Trichosporon loubieri from beef cattle[J]. , 2018, 30(1): 26-35.

参考文献

[1] 夏志宽,杨蓉娅. 毛孢子菌病[J]. 实用皮肤病学杂志,2010,3(4):215-217, 220.
XIA Z K, YANG R Y.Trichosprosis[J]. Practical Journal of Dermatology, 2010, 3(4):215-217, 220. (in Chinese with English abstract)
[2] 郝震锋, 敖俊红. 毛孢子菌病治疗研究进展[J]. 中国医师杂志,2004,6(1):141-142.
HAO Z F, AO J H.Research progress in the treatment of Trichosprosis[J]. Journal of Chinese Physician, 2004, 6(1):141-142. (in Chinese)
[3] 聂建巍. γ-干扰素增强RAW264.7巨噬细胞吞噬、杀伤阿萨希毛孢子菌的实验研究[D].合肥:安徽医科大学,2012.
NIE J W.Enhancement effect of IFN-γ on the phagocytosis, killing activity of RAW264.7 macrophages against Trichosporon asahii[D]. Hefei: Anhui Medical University, 2012. (in Chinese with English abstract)
[4] 郭莉娜,徐英春,窦红涛,等. 临床相关毛孢子菌感染的研究现状[J]. 中国真菌学杂志,2010,5(1):53-56.
GUO L N, XU Y C, DOU H T, et al.Research status in clinical infection caused by Trichosporon[J]. Chinese Journal of Mycology, 2010, 5(1):53-56. (in Chinese)
[5] WALSH T J, MELCHER G P, LEE J W, et al.Infections due to Trichosporon species: new concepts in mycology, pathogenesis, diagnosis and treatment[J]. Current Topics in Medical Mycology, 1993, 5:79-113.
[6] RISSI D R, KIRBY K D, SANCHEZ S.Systemic Trichosporon loubieri infection in a cat[J]. Journal of Veterinary Diagnostic Investigation, 2016, 28(3):350-353.
[7] 王端礼. 医学真菌学实验室检验指南[M]. 北京:人民卫生出版社,2005.
[8] 张悦天,马晓平,古玉,等.大熊猫源石膏样小孢子菌的分离鉴定与致病性研究[J]. 中国兽医科学,2015(6):551-559.
ZHANG Y T,MA X P,GU Y,et al.Identification and pathogenicity of Microsporum gypseum isolated from Ailuropoda melanoleuca[J]. Veterinary Science in China, 2015(6):551-559. (in Chinese with English abstract)
[9] 秦曼,向奇,马晓平,等.四川家兔源万博节皮菌的分离鉴定及其致病性研究[J]. 中国兽医科学,2016(9):1079-1086.
QIN M, XIANG Q, MA X P, et al.Identification and pathogenicity of Arthroderma vanbreuseghemii isolated from rabbit in Sichuan[J]. Veterinary Science in China, 2016(9):1079-1086. (in Chinese with English abstract)
[10] 宋秋荷,周会祥,赖雪花. 铜圈小培养鉴定真菌菌种的临床应用[J]. 九江学院学报(自然科学版),2013,28(1):76-77.
SONG Q H, ZHOU H X, LAI X H.The clinical application of small copper ring culture for the identification of fungi[J]. Journal of Jiujiang University (Natural Sciences), 2013, 28(1):76-77. (in Chinese)
[11] 马晓平,古玉,刘小敏,等. 大熊猫源枝孢样枝孢霉的分离鉴定[J]. 中国兽医科学,2013 (12):1217-1223.
MA X P, GU Y, LIU X M, et al.Isolation and identification of Cladosporium cladosporioides from Ailuropoda melanoleuc[J]. Veterinary Science in China, 2013 (12):1217-1223. (in Chinese with English abstract)
[12] 夏志宽,王文岭,杨蓉娅. 国内临床分离阿萨希毛孢子菌3个新基因型分析[J]. 中华医院感染学杂志,2012,22(14):2988-2990.
XIA Z K, WANG W L, YANG R Y.Three new genotypes of Trichosporon asahii strains isolated from Chinese patients[J]. Chinese Journal of Nosocomiology, 2012, 22(14):2988-2990. (in Chinese with English abstract)
[13] 曾秋林,陈灵敏,闵志刚. PAS染色显示真菌在临床上的应用[J]. 检验医学与临床,2010,7(21):2425.
ZENG Q L,CHEN L M,MIN Z G.The clinical application of PAS staining the fungi.[J]. Laboratory Medicine and Clinics, 2010, 7(21):2425. (in Chinese)
[14] 尹琦,陆贞妮,刘和艳,等. 5种抗真菌药物对常见感染酵母菌的抗菌活性分析[J]. 广西医科大学学报,2008,25(5):786-787.
YI Q, LU Z N, LIU H Y, et al.The antifungal activity analysis of five antifungal agents to common infection of yeast[J]. Journal of Guangxi Medical University, 2008, 25(5):786-787. (in Chinese)
[15] 马晓平, 杨秋霞, 李长城, 等. 发情期大熊猫阴道真菌的分离鉴定及酵母菌药敏试验[J]. 中国兽医科学, 2016(10):1233-1240.
MA X P, YANG Q X, LI C C, et al.Isolation and identification of fungi from the vagina of estrus giant panda and drug sensitive tests of the isolated yeasts[J]. Veterinary Science in China, 2016(10):1233-1240. (in Chinese with English abstract)
[16] BRYAN L K, PORTER B F, WICKES B L, et al.Meningoencephalitis in a dog due to Trichosporon montevideense[J]. Journal of Comparative Pathology, 2014, 151(2):157-161.
[17] GONZALEZ R N, WILSON D J, SICKLES S A, et al.Outbreaks of clinical mastitis caused by Trichosporon beigelii in dairy herds[J]. American Veterinary Medical Association Journal, 2001, 218(2):238-242.
[18] PADHYE A A, VERGHESE S, RAVICHANDRAN P, et al.Trichosporon loubieri infection in a patient with adult polycystic kidney disease[J]. Journal of Clinical Microbiology, 2003, 41(1):479-482.
[19] BHASKARAN A, OBEID K M, ARBEFEVILLE S, et al.Trichosporon loubieri fungemia in a 39-year-old caucasian woman with B-cell lymphoblastic leukemia[J]. Laboratory Medicine, 2016, 47(3):255-258.
[20] MARTY F M, BAROUCH D H, COAKLEY E P, et al.Disseminated trichosporonosis caused by Trichosporon loubieri[J]. Journal of Clinical Microbiology, 2003, 41(11):5317-5320.
[21] 夏志宽,赵逊,王文岭,等. rRNA基因序列分析在阿萨希毛孢子菌鉴定中的应用[J]. 中国真菌学杂志, 2008, 3(3):186-188.
XIA Z K, ZHAO X, WANG W L, et al.The application of rRNA gene sequence analysis in the identification of Trichosporon asahii[J]. Chinese Journal of Mycology, 2008, 3(3):186-188. (in Chinese)
[22] 张宇,郭良栋. 真菌DNA 条形码研究进展[J]. 菌物学报,2012,31(6):809-820.
ZHANG Y, GUO L D.Progress of fungal DNA barcode[J]. Mycosystema, 2012, 31(6):809-820. (in Chinese with English abstract)
[23] 曾莉莎,吕顺,郑芝波,等. 不同地区荷花腐败病菌生物学特性及IGS序列分析[J]. 热带作物学报,2017,38(1):136-143.
ZENG L S, LYU S, ZHENG Z B, et al.Biological characteristics and IGS sequence analysis of pathogen strains causing lotus rhizome rot(Fusarium commune) from different regions[J]. Chinese Journal of Tropical Crops, 2017, 38(1):136-143. (in Chinese with English abstract)
[24] 郭莉娜,徐英春,窦红涛,等.临床相关毛孢子菌的鉴定及体外药物敏感性研究[J]. 中国真菌学杂志,2009(2):82-86.
GUO L N,XU Y C,DOU H T, et al.Identification and in vitro antifungal susceptibility of medically important Trichosporon species[J]. Chinese Journal of Mycology, 2009(2):82-86. (in Chinese with English abstract)
[25] 李厚敏,刘伟,万哲,等. 临床相关毛孢子菌生物学特性的研究[J]. 中华检验医学杂志,2005(6):613-616.
LI H M,LIU W,WAN Z, et al.Microbiological characteristics of medically important Trichosporon species[J]. Chinese Journal of Laboratory Medicine, 2005(6):613-616. (in Chinese)
[26] MORETTI-BRANCHINI M L, FUKUSHIMA K, SCHREIBER A Z, et al. Trichosporon species infection in bone marrow transplanted patients[J]. Diagnostic Microbiology and Infectious Disease, 2001, 39(3):161-164.
[27] TAWARA S,IKEDA F,MAKI Ket al.In vitro activities of a new lipopeptide antifungal agent, FK463, against a variety of clinically important fungi[J]. Antimicrobial Agents and Chemotherapy, 1999, 44(1):57-62.
[28] 吕雪莲. 人类致病相关毛孢子菌表型和种系发生特征及种间鉴定方法的比较研究[D]. 北京:中国协和医科大学,2009.
LYU X L.A comparative study on the phenotypic and phylogenetic characteristics of human pathogenic related species and the identification methods of the species [D]. Beijing: Peking Union Medical College, 2009. (in Chinese)

肉牛源毛孢子菌Trichosporon loubieri的分离鉴定与药敏试验

Isolation, identification and drug sensitive test of Trichosporon loubieri from beef cattle

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	黄莉萍, 安莉丽, 李芳, 杨成年, 李虹, 吕光俊, 向枭, 孙翰昌, 翟旭亮, 朱成科. 中华鳖弗氏柠檬酸杆菌的鉴定及病理组织观察[J]. 浙江农业学报, 2020, 32(7): 1176-1186.
[2]	胡宗文, 杨娟, 苗春辉, 黄新球, 杨爽. 侵染狄斯瓦螨后西方蜜蜂的致畸等级划分[J]. 浙江农业学报, 2020, 32(7): 1187-1195.
[3]	陈章, 吴华健, 毛天骄, 韩业芹, 孙裴, 魏建忠, 李东风, 李郁. 猪链球菌2型制苗用菌株的筛选[J]. 浙江农业学报, 2020, 32(1): 57-64.
[4]	甯媛, 段琛, 常琼琼, 侯晓晖. 库蠓属6亚属翅形变化分析与亲缘关系[J]. 浙江农业学报, 2020, 32(1): 108-114.
[5]	严文一, 谢永东, 仰路希, 陈延, 王海霞, 孙勃, 贺忠群. 两个品种人参菜形态、解剖结构比较与核型分析[J]. 浙江农业学报, 2019, 31(8): 1321-1330.
[6]	林鸿煜, 钱晶, 严力蛟, 黄绍荣. 基于形态学空间格局分析与CA-Markov模型的武义县绿色基础设施时空格局变化及情景模拟[J]. 浙江农业学报, 2019, 31(7): 1193-1204.
[7]	张岩, 亓玉华, 鲁燕华, 杨乾坤, 何雨娟, 李俊敏, 陈剑平. 小麦黄花叶病毒P3蛋白致病功能域的鉴定和分析[J]. 浙江农业学报, 2019, 31(5): 777-783.
[8]	周会明, 张焱珍, 柴红梅, 杨荣情, 谭莹, 张萍萍, 白玉英, 赵一莲, 金玉洁. 一株临沧野生黄鸡?的鉴定与生长条件研究[J]. 浙江农业学报, 2019, 31(10): 1655-1662.
[9]	吴琼娟, 杨志鹏, 姚焱彬, 陆萍, 魏建忠, 孙裴, 李郁. 猪红斑丹毒丝菌制苗用菌株的筛选[J]. 浙江农业学报, 2018, 30(9): 1467-1475.
[10]	钟永军, 何昕蔚, 余达勇, 蒋晶晶, 江景勇. 台州猕猴桃溃疡病病原菌分子鉴定及药剂筛选[J]. 浙江农业学报, 2018, 30(9): 1548-1554.
[11]	袁雪梅, 姚嘉赟, 蔺凌云, 潘晓艺, 徐洋, 尹文林, 沈锦玉. 泥鳅致病性维氏气单胞菌的分离与鉴定[J]. 浙江农业学报, 2018, 30(5): 730-737.
[12]	王婧, 刁小龙, 杨茂义, 王全方, 陈晓兰, 张龙. 致仔猪腹泻大肠埃希菌的分离及耐药性分析[J]. 浙江农业学报, 2018, 30(4): 568-575.
[13]	刘子明, 金晶, 胡则辉, 戴庆敏, 李巧玲, 施倩, 吕耀平. 棘胸蛙白内障病原鉴定及病理组织观察[J]. 浙江农业学报, 2018, 30(11): 1811-1818.
[14]	朱凝瑜, 郑晓叶, 曹飞飞, 李杨阳, 郑天伦. 鳖源嗜水气单胞菌的耐药性研究[J]. 浙江农业学报, 2017, 29(9): 1445-1450.
[15]	吴庆岗, 张卫国, 常化文, 金保华, 刘朝霞. 基于梯度Hough变换的遮挡苹果目标定位[J]. 浙江农业学报, 2017, 29(6): 1009-1016.